LOCUS       sequence-240870-        6555 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses shRNA targeting ATRX. Construct has inverted repeats to be
            used in Sleeping beauty system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6555)
  AUTHORS   Nunez FJ, Mendez FM, Kadiyala P, Alghamri MS, Savelieff MG, 
            Garcia-Fabiani MB, Haase S, Koschmann C, Calinescu AA, Kamran N, 
            Saxena M, Patel R, Carney S, Guo MZ, Edwards M, Ljungman M, Qin T, 
            Sartor MA, Tagett R, Venneti S, Brosnan-Cashman J, Meeker A, 
            Gorbunova V, Zhao L, Kremer DM, Zhang L, Lyssiotis CA, Jones L, 
            Herting CJ, Ross JL, Hambardzumyan D, Hervey-Jumper S, Figueroa ME, 
            Lowenstein PR, Castro MG
  TITLE     IDH1-R132H acts as a tumor suppressor in glioma via epigenetic 
            up-regulation of the DNA damage response.
  JOURNAL   Sci Transl Med. 2019 Feb 13;11(479). pii: 11/479/eaaq1427. doi: 
            10.1126/scitranslmed.aaq1427.
  PUBMED    30760578
REFERENCE   2  (bases 1 to 6555)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Transl 
            Med."; date: "2019-02-13"; volume: "11(479). pii: 11/479/eaaq1427. 
            doi"; pages: " 10.1126/scitranslmed.aaq1427"
COMMENT     Sequence Label: pT2-shATRX
FEATURES             Location/Qualifiers
     source          1..6555
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     LTR             46..560
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    722..738
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(746..752,753..770,771..776))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    791..812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1271..1291)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1271..1289)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1310..1326)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1310..1326)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1334..1350
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1358..1364,1365..1382,1383..1388))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1403..1424
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1541..1558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1712..2300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1792..1811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2471..3262,3263..3331))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3094..3113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3332..3436)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3463..3918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3550..3569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3760..3781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4044..4066
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4058..4075
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4059..4075
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4085..4104
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4085..4103
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             4490..5006
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    5070..5411
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             5478..5894
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5824..5846
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5862..5878
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     ncRNA           5934..6055
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           6083..6097
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           6125..6251
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     primer_bind     complement(6284..6304)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     6531..6550
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
       61 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac
      121 tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa
      181 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
      241 gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
      301 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
      361 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc
      421 cagtcctccg atagactgcg tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
      481 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
      541 cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt tggagaacaa cattctgagg
      601 gtaggagtcg aatattaagt aatcctgact caattagcca ctgttttgaa tccacatact
      661 ccaatactcc tgaaatagtt cattatggac agcgcagaaa gagctgggga gaattgtgaa
      721 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
      781 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
      841 agtcgggaaa cctgtcgtgc cagatccatg gacttcacta gtgcgcgcgg ccgctctaga
      901 tggccagatc tagcttgtgg aaggctactc gaaatgtttg acccaagtta aacaatttaa
      961 aggcaatgct accaaatact aattgagtgt atgtaaactt ctgacccact gggaatgtga
     1021 tgaaagaaat aaaagctgaa atgaatcatt ctctctacta ttattctgat atttcacatt
     1081 cttaaaataa agtggtgatc ctaactgacc taagacaggg aatttttact aggattaaat
     1141 gtcaggaatt gtgaaaaagt gagtttaaat gtatttggct aaggtgtatg taaacttccg
     1201 acttcaactg tatagggatc ctctagctag agtcgacctc gcgggggggc ccggtaccca
     1261 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     1321 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1381 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1441 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     1501 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1561 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1621 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1681 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1741 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1801 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1861 gatacctgtc cgcctttctc ccttcgggaa ccgtggcgct ttctcatagc tcacgctgta
     1921 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1981 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2041 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2101 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2161 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2221 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2281 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2341 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2401 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2461 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2521 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2581 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2641 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2701 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2761 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2821 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2881 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaac ttggccgcag
     2941 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3001 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3061 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3121 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3181 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3241 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3301 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3361 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3421 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     3481 cattaagcgc ggcgggagtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3541 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3601 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3661 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3721 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3781 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3841 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3901 tattaacgct tacaatttcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     3961 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     4021 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg
     4081 cgcgtaatac gactcactat agggcgaatt ggagctcgga tccctataca gttgaagtcg
     4141 gaagtttaca tacacttaag ttggagtcat taaaactcgt ttttcaacta ctccacaaat
     4201 ttcttgttaa caaacaatag ttttggcaag tcagttagga catctacttt gtgcatgaca
     4261 caagtcattt ttccaacaat tgtttacaga cagattattt cacttataat tcactgtatc
     4321 acaattccag tgggtcagaa gtttacatac actaagttga ctgtgccttt aaacagcttg
     4381 gaaaattcca gaaaatgatg tcatggcttt agaagcttga tctgaagcct atagagtacg
     4441 agccatagat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     4501 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac
     4561 ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg
     4621 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     4681 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     4741 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     4801 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     4861 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt
     4921 gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc agattgattg
     4981 actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag acccctgccc
     5041 agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc gtgtctgtct
     5101 ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg tactagttag
     5161 ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga acacccggcc
     5221 gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg acctgaggaa
     5281 gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga gacgagaacc
     5341 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga agccgcgcgt
     5401 cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt
     5461 atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact
     5521 ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg
     5581 ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct
     5641 ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac
     5701 acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct
     5761 gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc
     5821 tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc
     5881 cttctctagg cgccggaatt agatctctcg actagggata acagggtaat tgtttgaatg
     5941 aggcttcagt actttacaga atcgttgcct gcacatcttg gaaacacttg ctgggattac
     6001 ttcttcaggt taacccaaca gaaggctcga gaaggtatat tgctgttgac agtgagcgcc
     6061 cgcttggatg gttctactaa tagtgaagcc acagatgtat tagtagaacc atccaagcgg
     6121 ttgcctactg cctcggaatt caaggggcta ctttaggagc aattatcttg tttactaaaa
     6181 ctgaatacct tgctatctct ttgatacatt tttacaaagc tgaattaaaa tggtataaat
     6241 taaatcactt ttttcaattc gccagcacag tggtcgaccc tgtggaatgt gtgtcagtta
     6301 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6361 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6421 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     6481 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6541 gaggccgagg ccgcc
//
